This application is related to PCT publication WO2007019266, entitled “KILLING HUMAN LYMPHOMA AND LEUKEMIA CANCER CELLS AND TCR-ACTIVATED NORMAL HUMAN CELLS BY DOPAMINE D1R AGONISTS” (the inventor of which, Dr. Mia Levite, being one of the present inventors), the entire contents of which are hereby incorporated herein by reference. This PCT publication discloses the unexpected activity of fenoldopam mesylate as a drug that potently and rapidly kills cancer cells, and specifically various types of human leukemia and lymphoma of T-cell or B-cell or non-T non-B origin, that express the D1R on their cell surface, thereby revealing the unknown ability of fenoldopam mesylate to act as an anti-cancer agent.
In PCT publication WO2007019266, the following findings are described:
1) Some types of human and mouse lymphoma (among them several types of T-cell lymphoma and leukemia (among them T-cell leukemia)) have moderate-dramatic elevation in the levels of dopamine D1 receptors expressed on their cell surface, in contrast to normal (i.e., non cancerous) human resting peripheral T-cells, which express negligible levels, if at all, of D1 dopamine receptors.
2) Exposing in vitro five different types of T-cell, B-cell or myeloid type human lymphoma and leukemia lines (specifically, the T-cell leukemia line, Jurkat; the human T-lymphoma line, the HuT-78 cutaneous ‘Sezary’ T-lymphoma; the human Chronic-Myeloid Leukemia (CML) line, K-562; and the human B-lymphoma lines: Burkitt's lymphoma lines Daudi and Raji) to concentration range of 1 mM-0.01 mM of fenoldopam mesylate or to several concentrations of other highly selective D1R agonists, leads to the death of all, or the vast majority of, these cancer cells.
The selective dopamine D1/5 receptor agonists tested and found effective in killing lymphoma and leukemia are: (1R-cis)-1-(aminomethyl)-3,4-dihydro-3-tricyclo[3.3.1.13,7]dec-1-yl-[1H]-2-benzopyran-5,6-diol hydrochloride (TOCRIS Cookson Product name: A 77636 hydrochloride; Catalog number: 1701; referred to as “potent, selective D1-like agonist; orally active”), (±)-1-phenyl-2,3,4,5-tetrahydro-(1H)-3-benzazepine-7,8-diol hydrobromide (TOCRIS COOKSON Product name: SKF 38393 hydrobromide; Catalog number: 0922; referred to as “D1-like dopamine receptor selective partial agonist”), and cis-(±)-1-(aminomethyl)-3,4-dihydro-3-phenyl-1H-2-benzopyran-5,6-diol hydrochloride (TOCRIS COOKSON Product name: 1534; Catalog number: A 68930 hydrochloride; referred to as “potent and selective D1-like dopamine receptor agonist”).
3) The killing of lymphoma and leukemia by fenoldopam mesylate and all the other selective dopamine D1/5 receptor agonists is dose dependent. Nevertheless, as expected, some D1R agonists were much more effective than others, and could kill the cancer cells in lower concentrations than the others. Fenoldopam mesylate and A 77636 hydrochloride were the most effective cancer killers and are thus preferred embodiments for use in the present invention.
4) Cell death induced by the fenoldopam mesylate or other highly specific dopamine D1R agonists is rapid (starting and measurable within 1-60 minutes after exposure), and occurred primarily via a mechanism of necrosis.
5) Most of the lymphoma and leukemia cells tested expressed on their cell surface markedly elevated levels not only of the D1R, but also of the dopamine D3 and dopamine D2 receptors, compared to much lower cell surface expression of the respective receptors on normal (not cancer) human T-cells. Yet, dopamine D2 and D3 receptor agonists exhibited much lower anti-cancer killing activity, if at all, compared to the effect exerted by the dopamine D1R agonists.
In PCT publication WO2007019266, it is further claimed that the findings described therein are relevant to novel treatment not only of cancers that express the D1R, but also to the treatment of diseases caused by activated T-cells, among them being, for example, T-cell mediated autoimmune diseases, T-cell mediated inflammatory diseases, T-cell mediated graft rejection and T-cell mediated Graft Versus Host Diseases (GVHD), the two latter occurring after mis-matched organ or cell transplantation. This is claimed based in the following findings also described in PCT publication WO2007019266:
A. T-cell receptor (TCR)-activated normal (i.e., non cancerous) human peripheral T-cells express dramatically elevated levels of dopamine D1 receptors on their cell surface, as opposed to resting normal human peripheral T-cells that do not express this receptor, or do so to minimal not significant levels.
B. Exposing TCR-activated human normal peripheral T-cells in vitro to several highly selective dopamine D1R agonists, such as fenaldopam mesylate, kills potently a substantial proportion of these activated T-cells, while hardly affecting the resting (i.e., not TCR-activated) human normal peripheral T-cells. The killing of TCR-activated T-cells by all the selective dopamine D1R agonists is dose dependent, as seen for the killing of cancer cells, and also, here, as expected, some D1R agonists are much more effective than others, and could kill the cancer cells in lower concentrations than the others. Of all the highly selective D1R agonists tested therein, fenoldopam mesylate and A 77636 hydrochloride were the most effective killers of TCR-activated T-cells.
Thus, the findings described in PCT publication WO2007019266 reveal the previously unknown ability of fenoldopam mesylate to kill not only cancer cells of T, B and non-T non-B origin (CML being an example of the latter type), that express D1R, but also to kill non-cancerous normal (yet potentially causing other diseases than cancer) TCR-activated human T-cells, while causing only minimal death of resting normal human T-cells. These findings suggest that fenoldopam can potentially act not only as a drug for D1R expressing cancers, but also as a drug for T-cell mediated (or T-cell associated) autoimmune diseases, as in such diseases the damage is caused by TCR-activated T-cells that are directed towards a specific self antigen and are activated by such autoantigen over and over again.
The following is a list of all accepted autoimmune diseases, each of which is expected to be treatable by means of the present invention: acute disseminated encephalomyelitis (ADEM), Addison's disease, ankylosing spondylitis, antiphospholipid antibody syndrome (APS), aplastic anemia, autoimmune hepatitis, autoimmune oophoritis, coeliac disease, Crohn's disease, diabetes mellitus type 1, gestational pemphigoid, Goodpasture's syndrome, Graves' disease, Guillain-Barré syndrome, Hashimoto's disease, idiopathic thrombocytopenic purpura, Kawasaki's disease, lupus erythematosus, multiple sclerosis, myasthenia gravis, opsoclonus myoclonus syndrome (OMS), optic neuritis, Ord's thyroiditis, pemphigus, pernicious anaemia, polyarthritis in dogs, primary biliary cirrhosis, rheumatoid arthritis, Reiter's syndrome, Sjögren's syndrome, Takayasu's arteritis, temporal arteritis (also known as “giant cell arteritis”), warm autoimmune hemolytic anemia, and Wegener's granulamatosis. The following is a list of diseases suspected or theorized to be linked to autoimmunity, each of which are expected to be treatable by means of the present invention: alopecia universalis, Behçet's disease, Chagas' disease, chronic fatigue syndrome, dysautonomia, endometriosis, hidradenitis suppurativa, interstitial cystitis, Lyme disease, morphea, neuromyotonia, narcolepsy, psoriasis, sarcoidosis, schizophrenia, scleroderma, ulcerative colitis, vitiligo, and vulvodynia.
In general, the autoimmune diseases can be broadly divided into systemic and organ-specific or localized autoimmune disorders, depending on the principal clinico-pathologic features of each disease.
A partial list of human T-cell mediated autoimmune diseases, each of which is expected to be treatable by means of the present invention, includes the following: insulin-dependent (type 1) diabetes mellitus, multiple sclerosis, myasthenia gravis, autoimmune myocarditis, alopecia aerate (the latter now understood to represent an organ-restricted, T cell-mediated autoimmune disease of hair follicles), and others.
Fenoldopam mesylate was also suggested in PCT publication WO2007019266 to act against anti-inflammatory diseases, or against any other disease in which the detrimental cells express the D1R, and whereby the binding of this receptor by fenoldopam mesylate kills these cells.
The inflammatory diseases are disease characterized by inflammation. These diseases, each of which is expected to be treatable by means of the present invention, are listed in: www.thefreedictionary.com/inflammatory+disease.
TCR-activated T-cells are also detrimental in cases of organ transplantation, as they are the cells that recognize the transplanted organ as foreign and reject it, as well as in cases of GVHD, as they are the transplanted cells that recognize the new body as foreign, and attack it.
Based on the ability of fenoldopam to eradicate TCR-activated T-cells, described in PCT publication WO2007019266, while sparing resting T-cells, it is suggested therein to potentially act also as an anti-graft rejection drug, and thus to prevent the rejection of transplanted organs or cells, as well as an anti-GVHD drug, and thus prevent the devastating consequences of this disease, which often kills the transplanted patient.
Fenoldopam mesylate has a very short half life: T1/2=˜5 min, and is rapidly eliminated upon discontinuation of the IV infusion.
While fenoldopam mesylate is currently used in the clinic in an FDA-approved protocol of a single 24 hr-48 hr IV administration for acute hypertension, other new formulations are needed to get an optimal effect for its new indications, including anti-cancer, anti-autoimmune, anti-inflammatory, anti-graft rejection, and anti-GVHD agent. For all these new indications, it may be of high advantage to have in hand a drug formulation that can be given at multiple times, or that is released slowly and continuously over periods from a few hours to a few weeks or months, or with a longer half life in the body, or that can be injected locally into the tumor or into autoimmune-afflicted organs and released therein over a desired period in a regulated manner, and/or suitable for treatment of a chronic pathological condition (rather than acute one) or other.
Fenoldopam mesylate (6-chloro-2,3,4,5-tetrahydro-1-(4-hydroxyphenyl)-1H-3-benzazepine-7,8-diol, methanesulfonate) is a highly selective agonist for the dopamine D1 receptor (termed D1R receptor or dopamine DA1 agonist), causing an increased splanchnic and renal blood flow with a strong diuretic and natriuretic effect. It possesses several decisive advantages as an antihypertensive and for the treatment of congestive heart failure over commonly used other drugs.
Fenoldopam mesylate is used in the clinic for its vasodilatory actions mainly in the treatment of severe hypertension. It has only peripheral actions and does not cross the blood-brain barrier (BBB). The major side effects reported in 10% (or less) of the patients are headache, flushing and nausea
The solubility of fenoldopam mesylate is 9.1 mg/ml in an HCl/KCl buffer pH 1.5, but decreases to 0.13 mg/ml in buffer pH 6.8 and to 0.06 mg/ml in pH 7.5. Free base fenoldopam is hydrophobic and can be incorporated in a formulation for improved drug load and release profile. Fenoldopam is a white to off-white powder with a molecular weight of 401.87 and a molecular formula of C16H16ClNO3.CH3SO3H. It is sparingly soluble in water, ethanol and methanol, and is soluble in propylene glycol. Ampules for injection contain in each 1 mL in sterile aqueous solution, citric acid 3.44 mg; fenoldopam mesylate equivalent to fenoldopam 10 mg; propylene glycol 518 mg; sodium citrate dihydrate 0.61 mg; sodium metabisulfite 1 mg (Formula 1).

This infusion is given only at acute situations to treat blood pressure disorders or problems of patients, primarily those suffering from kidney disorders or problems.
There are no other clinical formulations for this drug, not an oral or even a non-infused parenteral formulation. There are a few patents describing tablet type formulations for oral use and transdermal delivery. U.S. Pat. Nos. 6,238,693 and 6,960,353 describe compositions for application to skin to administer fenoldopam by permeation through the skin, using a permeation enhancer and hydrogels. The disclosed transdermal application of fenoldopam is for the treatment of hypertension, congestive heart failure, and chronic and acute renal failure. Moreover, these formulations did not address formulation issues such as limited solubility of fenoldopam which requires the development of specific formulations for poorly soluble drugs. Moreover, this drug possesses ortho-resorcinol groups which are highly sensitive to oxidation, and with limited stability in general. Also, formulations for extended release were not described for this drug, other than one publication which describes a coated tablet for oral administration releasing fenoldopam and succinic acid for a few hours in vitro (European Journal of Pharmaceutics and Biopharmaceutics, 46:105-113 (1998)).